华支睾吸虫成虫抗原和排泄分泌产物对T细胞的作用研究

发布时间：2018-04-04 05:25

  本文选题：华支睾吸虫　切入点：DC细胞　出处：《中国人兽共患病学报》2017年06期

【摘要】：目的观察华支睾吸虫成虫抗原(Crude antigen,CA)、排泄/分泌产物(excretory-secretory products,ESPs)对T细胞的作用。方法体外分离小鼠骨髓细胞诱导分化为未成熟骨髓树突状细胞(immature DC,iDC);磁珠分选仪分选小鼠脾脏细胞的初始CD4+T细胞;流式细胞术检测DC、CD4+T细胞的纯度;抗原刺激DC细胞,实验分为PBS阴性对照组,LPS阳性对照组,CA和ESPs刺激组;负载抗原后的DC细胞与分选的CD4+T细胞共培养72h;Real time-PCR检测T-bet、GATA3mRNA的相对表达量;ELISA检测细胞培养上清中IFN-γ、IL-4细胞因子的表达量。结果与PBS组相比,ESPs刺激组Tbet、GATA3mRNA表达水平升高(P0.05),而CA刺激组T-bet、GATA3 mRNA表达水平差异不显著;与PBS组相比,ESPs刺激组细胞因子IFN-γ、IL-4的含量均升高(P0.05),CA刺激组仅IFN-γ的分泌增高(P0.05),IL-4无明显变化。结论 CA可能诱导宿主产生Th1型免疫应答,ESPs可能诱导宿主产生Th1型、Th2型免疫应答。
[Abstract]:Objective to observe the effects of the antigens of Clonorchis sinensis (Crude antigenia CAA), excretory-secretory products (ESPs) on T cells.Methods Murine bone marrow cells were isolated and differentiated into immature bone marrow dendritic cells in vitro. The initial CD4 T cells were isolated from mouse spleen cells by magnetic bead sorter, the purity of CD4 T cells was detected by flow cytometry, and DC cells were stimulated by antigen.The experiment was divided into PBS negative control group, LPS-positive control group, CA and ESPs stimulation group, DC cells loaded with antigen were co-cultured with CD4 T cells for 72 h, Real time-PCR was used to detect the relative expression of T-bettgata-GATA3 mRNA and Elisa was used to detect the expression of IFN- 纬 IL-4 cytokines in the supernatant of cell culture.缁撴灉涓嶱BS缁勭浉姣，

本文编号：1708559